Stereoselectivity of Microbial Lipases

Stadler, Peter; Kovac, Andrea; Haalck, Lutz; Spener, Fritz; Paltauf, Fritz

doi:10.1111/j.1432-1033.1995.tb20394.x

Cited by 64 publications

(41 citation statements)

References 46 publications

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“…Consequently, either the carbonyl oxygen (O7) of the compound or the side chains of the appropriate residues must reorient to prevent unfavourable proteinϪligand interactions. before for other lipases [38,40]. the HA pocket, where the sn-3 fatty acid chain binds (Fig.…”

Section: Resultsmentioning

confidence: 99%

Structural basis of the chiral selectivity of Pseudomonas cepacia lipase

Lang

Mannesse

Haas

et al. 1998

European Journal of Biochemistry

141

102

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To investigate the enantioselectivity of Pseudomonas cepacia lipase, inhibition studies were performed with S C -and R C -(R P ,S P )-1,2-dialkylcarbamoylglycero-3-O-p-nitrophenyl alkylphosphonates of different alkyl chain lengths. P. cepacia lipase was most rapidly inactivated by R C -(R P ,S P )-1,2-dioctylcarbamoylglycero-3-O-p-nitrophenyl octylphosphonate (R C-trioctyl) with an inactivation half-time of 75 min, while that for the S C -(R P ,S P )-1,2-dioctylcarbamoylglycero-3-O-p-nitrophenyl octyl-phosphonate (S C -trioctyl) compound was 530 min. X-ray structures were obtained of P. cepacia lipase after reaction with R Ctrioctyl to 0.29-nm resolution at pH 4 and covalently modified with R C-(RP ,SP)-1,2-dibutylcarbamoylglycero-3-O-p-nitrophenyl butyl-phosphonate (R C -tributyl) to 0.175-nm resolution at pH 8.5. The three-dimensional structures reveal that both triacylglycerol analogues had reacted with the active-site Ser87, forming a covalent complex. The bound phosphorus atom shows the same chirality (S P) in both complexes despite the use of a racemic (R P ,S P ) mixture at the phosphorus atom of the triacylglycerol analogues. In the structure of R C -tributyl-complexed P. cepacia lipase, the diacylglycerol moiety has been lost due to an aging reaction, and only the butyl phosphonate remains visible in the electron density. In the R C-trioctyl complex the complete inhibitor is clearly defined; it adopts a bent tuning fork conformation. Unambiguously, four binding pockets for the triacylglycerol could be detected : an oxyanion hole and three pockets which accommodate the sn-1, sn-2, and sn-3 fatty acid chains. Van der Waals' interactions are the main forces that keep the radyl groups of the triacylglycerol analogue in position and, in addition, a hydrogen bond to the carbonyl oxygen of the sn-2 chain contributes to fixing the position of the inhibitor.Keywords : crystal structure; transition-state analog; enantioselectivity; lipase; stereospecificity.Lipases are lipolytic enzymes, which hydrolyze ester bonds fold, a structural motif common to a wide variety of hydrolases of triacylglycerols. However, their substrate specificity is not [3]. Their active sites contain a catalytic triad, Ser-His-Asp/Glu, limited to triacylglycerols. They may also hydrolyze ester bonds similar to those of serine proteases [4Ϫ6]. Studies with lipases of other compounds such as acetyl-arylpropionic acid esters, covalently complexed with organosulfates [7], organophosphates which are precursors for the nonsteroidal anti-inflammatory [8, 9], or organophosphonates [10Ϫ12] demonstrated that, in the agents naproxen and ibuprofen [1]. Because of this broad sub-presence of lipid-like compounds or organic solvents, their strate specificity, and because of their distinct stereopreferences, active-site regions may undergo drastic conformational changes, lipases have found widespread application in the enantioselec-exposing the catalytic residues and the surrounding hydrophobic tive synthesis of organic compounds, and in the resolution of ...

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Section: Resultsmentioning

confidence: 99%

Structural basis of the chiral selectivity of Pseudomonas cepacia lipase

Lang

Mannesse

Haas

et al. 1998

European Journal of Biochemistry

141

102

View full text Add to dashboard Cite

show abstract

“…To systematically assess the influence of the sn-2 substituent on the steric course of hydrolysis, Stadler et al [46] determined the stereoselectivity of four microbial lipases [ROL, CRL, and the lipases from Pseudomonas glumae (PGL), and Pseudomonas sp. (PSL)] toward a series of triacylglycerol analogs with different polarities at sn-2 position of the glycerol backbone (acyloxy, alkyloxy, alkyl, or phenyl group) (Fig.…”

Section: Stereoselectivity Toward Triacylglycerol Analogsmentioning

confidence: 99%

“…Also nonspecific enzymes have been described, e.g. CRL [46][47][48]51] and porcine pancreatic lipase [43,49,50].…”

Section: Stereoselectivity Toward Prochiral Triacylglycerolsmentioning

confidence: 99%

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Molecular basis of lipase stereoselectivity

Kovac

Scheib

Pleiss

et al. 2000

Eur. J. Lipid Sci. Technol.

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“…In addition to the common capability for lipases to cleave ester bonds, some lipases hold special specificities, which are essentially important for EA isolation and enrichment (Piazza et al, 1992;Sonnet et al, 1993a;Stadler et al, 1995;Lee and Parkin, 2000).…”

Section: Lipase Specificities For Ea Enrichmentmentioning

confidence: 99%